Complete secretion of recombinant Bacillus subtilis levansucrase in Pichia pastoris for production of high molecular weight levan

文献类型： 外文期刊

第一作者： Chen, Shuochang

作者： Chen, Shuochang;Tong, Qiuping;Guo, Xiaolei;Cong, Hao;Zhao, Zi;Liang, Wenfeng;Zhu, Ping;Yang, Hui;Chen, Shuochang;Guo, Xiaolei;Zhao, Zi;Liang, Wenfeng;Yang, Hui;Chen, Shuochang;Guo, Xiaolei;Zhao, Zi;Liang, Wenfeng;Yang, Hui;Li, Jiemin

作者机构：

关键词： Thermostable levansucrase; Pichia pastoris; Complete secretion; Glycosylation; High molecular weight levan

期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.025； 五年影响因子：7.626 ）

ISSN： 0141-8130

年卷期： 2022 年 214 卷

页码：

收录情况： SCI

摘要： Three signal peptides from alpha-mating factor (alpha-MF), inulinase (INU) and native levansucrase (LS) were compared for secretion efficiency of Bacillus subtilis levansucrase SacB-T305A in Pichia pastoris GS115. The first complete secretion of bacterial levansucrase in yeasts under methanol induction was achieved while using alpha-MF signal. The secreted recombinant Lev(alpha-MF) proved to be glycosylated by combination of NanoLC-MS/MS and Endo H digestion. Interestingly, glycosylation not only improved significantly the polymerase thermostability, but also reversed the products profiles to favor synthesis of high molecular weight (HMW) levan which accounted for approximately 73 % to total levan-type polysaccharides. It indicated for the first time that the glycosylation of recombinant B. subtilis levansucrase affected significantly the products molecular weight distribution. It also provided a promising enzymatic way to effectively product HMW levan from sucrose resources.

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